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Nigrotectal projection to the inferior colliculus: horseradish peroxidase transport and tyrosine hydroxylase immunohistochemical studies in rats, cats, and bats.

The present study investigated descending projections from the substantia nigra to the auditory tectum. Small (0.02-0.05 microliters) injections of a 30-60% aqueous solution of horseradish peroxidase (HRP) were made unilaterally into the inferior colliculus in rats, cats, and bats (Eptesicus fuscus). Tissue blocks including the substantia nigra, superior colliculus, and inferior colliculus were removed, sectioned, and processed for visualization of HRP. Results show that the substantia nigra, pars lateralis, projects to the inferior colliculus ipsilaterally. In addition, retrogradely labeled cells are found dorsal to the pars lateralis, in a column within the lateral tegmental area of the midbrain. Analysis of injection sites suggests that the principal target of this nigral projection is the dorsal and rostral pericentral region of the inferior colliculus. Immunohistochemical studies with an antibody to tyrosine hydroxylase demonstrate catecholaminergic neurons within the pars lateralis and lateral tegmentum that are similar in location and morphology to one class of HRP retrogradely labeled cells within these structures. These immunohistochemical studies also demonstrate a plexus of fine, varicose tyrosine hydroxylase-positive axons in the rostral pericentral region of the colliculus. The presence of this nigrotectal projection to the inferior colliculus is discussed in relation to its possible role in the control of acousticomotor behavior.

Animals↗

Functional organization of ascending and descending connections of the cochlear nucleus of horseshoe bats.

The ascending projections of the cochlear nucleus (CN) and the sources of descending inputs to the CN were investigated in horseshoe bats (Rhinolophus rouxi) by tracing the anterograde and retrograde transport of horseradish peroxidase (HRP or WGA-HRP) injected into the CN. The tracer was iontophoretically deposited into physiologically characterized regions of the cochlear nucleus (Feng and Vater, '85). We report the course and termination of pathways arising from the anteroventral (AVCN), posteroventral (PVCN), and dorsal (DCN) cochlear nucleus. The projection fields within the auditory brainstem centers (superior olivary complex [SOC]; lateral lemniscus complex [LLC]; and inferior colliculus [IC]) and their tonotopic organization according to the frequency representations at the injection sites are described. While the projection pattern is generally in accordance with other mammals, several species-characteristic features are noted: i) the lateral superior olive (LSO) receives tonotopically organized input from both the AVCN and PVCN; ii) the CN-projections to medial nuclear groups of the SOC located between the LSO and the medial nucleus of the trapezoid body do not support previously suggested homologies; iii) the ventral nucleus of the LLC can be subdivided into two divisions with distinct input patterns from the AVCN and PVCN, respectively.

Animals↗

Calbindin-like immunoreactivity in the central auditory system of the mustached bat, Pteronotus parnelli.

With the aid of a polyclonal antibody specific for Calbindin D-28k, we studied the distribution of this calcium-binding protein in the central auditory system of the mustached bat, Pteronotus parnelli. Components of the cochlear nucleus (CN) that were calbindin-positive (cabp(+] included the root of the auditory nerve, multipolar and globular bushy cells in the anteroventral CN, multipolar and octopus cells in the posteroventral CN, and small and medium-size cells in the dorsal CN. Not stained were spherical bushy cells of the anteroventral CN and pyramidal/fusiform cells in the dorsal CN. In the superior olivary complex, labeled cells were found in the lateral and medial nuclei of the trapezoid body, the ventral and ventromedial periolivary nuclei, and the anterolateral periolivary nucleus. No cellular labeling was seen in the lateral superior olive. In the medial superior olive, only marginal cells were cabp(+). Labeled fibers could be seen surrounding the gosts of unlabeled cells in both the latter nuclei. Most cells in the intermediate nucleus and the columnar division of the ventral nucleus of the lateral lemniscus were cabp(+). However, the dorsal nucleus was cabp(-). A group of cabp(+) cells was also seen in the paralemniscal zone. The inferior colliculus had a relatively low density of cabp(+) cells. Labeled cells were more common in the caudal half of the central nucleus, and in the external nucleus and dorsal cortex. In the auditory thalamus, nearly every cell in the medial geniculate body was cabp(+), but those in the suprageniculate nucleus and in the posterior group did not stain. Small cells in the intermediate layer and giant cells in the deep layers of the superior colliculus were densely cabp(+). In the pons, cabp(+) cells and neuropil could be seen in the medial and lateral pontine nuclei (pontine gray). In conclusion, calbindin-like immunoreactivity was found in most of the brainstem auditory system, as well as in regions associated with acoustic orientation or control of vocalization. However, except for a minority of cells of the medial superior olive, it is conspicuously absent from the nuclei receiving binaural input below the level of the inferior colliculus.

Animals↗

Neuropeptide Y-like immunoreactivity in the diencephalon of the little brown bat (Myotis lucifugus): localized variations with physiological state.

The current study used light microscopic immunocytochemistry to demonstrate and compare neuropeptide Y-like immunoreactivity (NPY-IR) in the diencephalon of the little brown bat (Myotis lucifugus) at different stages in its annual cycle of activity and hibernation. Animals were sacrificed in each of three discrete physiological states: euthermic, hypothermic, and hibernating. In general, NPY-IR was abundant in the hypothalamus and sparse in other diencephalic areas. Immunoreactivity was present in a number of pathways which project to or originate from diencephalic nuclei; these include the ansa peduncularis, medial forebrain bundle, inferior thalamic peduncle, stria terminalis, stria medullaris, mammillary peduncle, and dorsal longitudinal fasiculus. Dense fiber plexuses were present throughout the hypothalamus; however, NPY-IR was conspicuously absent from the suprachiasmatic nucleus. Immunoreactive perikarya were located in the supraoptic, dorsomedial, ventromedial, and arcuate nuclei, in the external division of the ventral lateral geniculate nucleus, and in the pineal gland. Localized changes in density and/or distribution of NPY-IR were correlated with changes in physiological state.

Animals↗

Ultrastructure of the horseshoe bat's organ of Corti. II. Transmission electron microscopy.

The fine structure of the organ of Corti was investigated in the echolocating horseshoe bat (Rhinolophus rouxi) by transmission electron microscopy. Particular emphasis was placed on the receptor cells and their supporting cells. The receptor cells, inner hair cells (IHC) and outer hair cells (OHC), possess the typical mammalian shape, but OHCs are extremely short (length: 12-15 microns in the basal turn and up to 28-30 microns in the apical turn). The afferent innervation of both types of receptor cells and the efferent innervation of the IHC system conform to the general mammalian scheme; however, confirming earlier reports, an efferent innervation to the OHCs is absent. Throughout the cochlea, IHCs and OHCs possess a single layer of subsurface cisternae. Above the level of the nucleus of the OHCs, the arrangements of the subsurface cisternae and their connection to the lateral cell membrane via pillars are highly regular, whereas in IHCs, the cisternae are of irregular shape and the pillar system is much less distinct. In the basal turn of the cochlea, the attachment sites of the OHCs to the supporting cells possess specialized features: (a) in the reticular lamina, the contact sites of the cuticular plates of OHCs with the outer pillar cells and the Deiters cell phalanges are of exaggerated length, and (b) the cup formation of the Deiters cell body, which houses the bottom of the OHC, has a specialized shape and is packed with electron-dense material and microtubules. The results are discussed in relation to cochlear ultrastructure in other mammals and in the context of active processes in cochlear mechanics.

Afferent Pathways↗

Patterns of GABAergic immunoreactivity define subdivisions of the mustached bat's medial geniculate body.

The anatomy and the spatial distribution of neurons and axonal endings (puncta) immunoreactive for glutamic acid decarboxylase (GAD) or gamma-aminobutyric acid (GABA) were studied in the medial geniculate body of the mustached bat (Pteronotus parnellii). The principal findings are that: 1) most GABAergic neurons are present in the dorsal and ventral divisions with few, if any, in the medial division; 2) only a small fraction, about 1% or less, of auditory thalamic neurons are immunopositive; 3) the density of immunoreactive puncta is independent on the number of GABAergic neurons in the thalamic divisions, with the ventral division having the largest number/unit area, the medial division about 75% of this value, and the dorsal division only about 50%; and 4) the form of the puncta was unique to each division, those in the ventral division being medium-sized and comparatively simple, those in the medial division predominantly large, coarse, and complex, while dorsal division ending were finer and more delicate. These patterns recapitulate, with some significant exceptions, those found in the rat and cat. The puncta could originate from several sources; while many may arise from intrinsic GABAergic Golgi type II local circuit neurons, these cells may not be the only or even the principal source. Thus, the dorsal division contains comparatively many immunopositive cells though fewer puncta than might be expected if the bulk of these were to arise from auditory thalamic interneurons. This suggests that other, extrinsic sources, such as the thalamic reticular nucleus, may be the source of such endings. A second point is that the form and density of the puncta is regionally specific within the medial geniculate complex. These local patterns might have a significant and regionally specific role in controlling the differential excitability of auditory thalamic neurons. The distribution of presumptive synaptic endings also has implications for the number and arrangement of glomeruli or synaptic nests. Thus, these circuit elements, which are common to the thalamic nuclei in other species, might play an important role in local synaptic circuits between different types of cells. If so, then the structural variations embodied in these patterns could subserve functional arrangements that differ among species. Such patterns might reflect concomitant physiological differences in the organization of local circuits within the microchiropteran medial geniculate body.

Animals↗

Projections to the medial superior olive from the medial and lateral nuclei of the trapezoid body in rodents and bats.

In this study we present direct evidence of axonal projections from both the medial and lateral nuclei of the trapezoid body to the medial superior olive. Projections were traced by intracellularly labeling cells and axons in a tissue slice preparation of two rodent species, Mus musculus and Meriones unguiculatus and two bat species, Eptesicus fuscus and Pteronotus parnellii. The main axon of most principal cells in the medial nucleus of the trapezoid body gives off one or more collateral branches which arborize within the medial superior olive. These collateral axons form small bouton-like swellings which primarily contact somata within the central cell column in the medial superior olive. Likewise, labeled elongate and multipolar cells of the lateral nucleus of the trapezoid body send axons to both the medial and lateral superior olives. These axons also form perisomatic contacts in both target nuclei. These two sets of projections may relay ascending input to the medial superior olive and the lateral superior olive; the medial nucleus of the trapezoid body is known to relay input from the contralateral ventral cochlear nucleus, and the lateral nucleus of the trapezoid body may relay input from the ipsilateral ventral cochlear nucleus. These projections offer two routes for indirect, possibly inhibitory input to reach the medial superior olive from both cochlear nuclei. These indirect, inhibitory pathways may parallel the direct excitatory projections the medial superior olive receives from each cochlear nucleus.

Animals↗

The neurons of the medial geniculate body in the mustached bat (Pteronotus parnellii).

The neurons in the medial geniculate body were studied in Golgi preparations from adult mustached bats (Pteronotus parnellii). Their somatic and dendritic configurations were compared with those of cells in other, nonecholocating mammals. A second goal was to use the thalamic nuclear subdivisions derived from Golgi material to integrate the findings in parallel studies of cytoarchitecture, immunocytochemistry, and tectothalamic connections. Three primary divisions are defined. The ventral division is large and has a stereotyped neuronal organization. Medium-sized perikarya (about 10 microns in diameter) represent tufted neurons; the fibrodendritic plexus forms laminae in the lateral part along which midbrain axons terminate. A smaller, possibly intrinsic, neuron with thin, sparse dendrites is rarely impregnated. Neurons in the larger, medial part, which represents frequencies of 60 kHz and higher, have more spherical dendritic fields; their branching pattern remains tufted, and the laminar organization was less evident. The dorsal division is about equal in size, and it has many nuclei and a corresponding neuronal diversity. These neurons are medium-sized except in the suprageniculate nucleus, where many cells are larger. Four dorsal division nuclei are recognized. Each has neurons with radiate or weakly tufted dendritic arbors. Superficial dorsal nucleus neurons are oriented from medial to lateral, imparting a slightly laminated appearance to the neuropil. A few smaller, stellate neurons with modest dendritic domains are present. Suprageniculate nucleus neurons have radiating dendritic fields that project spherically; they have fewer branches than dorsal nucleus neurons. The posterior limitans nucleus is dorsomedial to the suprageniculate nucleus; it has small neurons with long, sparsely branched dendrites. The rostral pole nucleus, included in the dorsal division on cytoarchitectonic grounds, had too few neurons impregnated to reveal its neuronal architecture. The medial division, the smallest of the main parts, is one nucleus with at least six types of cells, including the magnocellular, bushy tufted, disc-shaped, medium-sized multipolar, elongated, and small stellate neurons. There is no laminar arrangement. Many of the neurons resemble those in rodent, marsupial, carnivore, and primate auditory thalamic nuclei. Despite such morphological correspondences, functional differences, such as the evolution of combination sensitivity, suggest that structurally comparable auditory thalamic neurons may subserve diverse physiological representations.

Animals↗

Convergence and divergence of ascending binaural and monaural pathways from the superior olives of the mustached bat.

The lateral superior olive and medial superior olive give rise to pathways that terminate in the dorsal nucleus of the lateral lemniscus and central nucleus of the inferior colliculus. In most mammals, neurons in both the medial and lateral superior olives are binaural, but in the mustached bat most neurons in the medial superior olive are monaural. The aims of this study were to determine how the inputs to the medial superior olive contribute to its monaurality and to determine whether the ascending projections from the lateral and medial superior olives overlap or remain segregated at their targets. Injections of two different tracers were placed in tonotopically matched areas of the lateral and medial superior olives in the same animal. Retrograde transport from injections in the medial superior olive labeled spherical cells in the contralateral anteroventral cochlear nucleus and principal cells in the ipsilateral medial nucleus of the trapezoid body. Few cells were labeled in ipsilateral cochlear nucleus. Anterograde transport resulted in tonotopically specific distributions of label with the same laterality as in nonecholocating mammals. In the dorsal nucleus of the lateral lemniscus, label from the lateral and medial superior olives largely overlapped. In the inferior colliculus, label from the lateral and medial superior olives largely overlapped. In the inferior colliculus, label from the two sources overlapped in the high and low frequency ranges, but in the frequency range around 60 kHz, label from the medial superior olive extended more dorsally than that from the lateral superior olive. These results indicate that projections of the lateral and medial superior olives overlap extensively at their targets.

Animals↗

Origin of ascending projections to the nuclei of the lateral lemniscus in the big brown bat, Eptesicus fuscus.

The nuclei of the lateral lemniscus in the echolocating bat, Eptesicus fuscus, are large and highly differentiated. In each nucleus, different characteristic response properties predominate. To determine whether the dissimilar response properties are due in part to differential ascending input, we examined the retrograde transport from small deposits of horseradish peroxidase (HRP) or HRP conjugated with wheat germ agglutinin (WGA-HRP) in the nuclei of the lateral lemniscus. The intermediate nucleus (INLL) and the two divisions of the ventral nucleus (VNLL) receive almost exclusively monaural input from the anteroventral and posteroventral cochlear nuclei and from the medial nucleus of the trapezoid body. Lesser inputs originate in the lateral nucleus of the trapezoid body and the ventral periolivary area. Although the three monaural nuclei of the lateral lemniscus all receive input from the same set of nuclei, and from the same identified cell types in the cochlear nucleus, there is a difference in the relative proportions of input from these sources. The dorsal nucleus (DNLL) receives input mostly from binaural structures, the lateral and medial superior olives and the contralateral DNLL, with only a minor projection from the cochlear nucleus. The lateral and medial superior olives project bilaterally; the bilateral projection from the medial superior olive is unusual in that it is found in only a few mammalian species. The results show a segregated pattern of binaural projections to DNLL and monaural projections to INLL and VNLL that is consistent with the binaural response properties found in DNLL and the exclusively monaural response properties found in INLL and VNLL. The differences in response properties between monaural nuclei, however, are not due to input from different nuclei or cell types but may be influenced by differing magnitudes of the constituent ascending projections.

Afferent Pathways↗

Seasonal changes in circulating steroid concentration and their correlation with the ovarian activity in the female Indian sheath-tailed bat, Taphozous longimanus.

The relationship between ovarian activity and circulating steroid concentration was studied in the female sheath-tailed bat, Taphozous longimanus. T. longimanus breeds twice in rapid succession during the year at Varanasi, India. Ovarian recrudescence was observed during September, and antral follicles were first observed during the month of October. Circulating androstenedione concentration showed an increase beginning in October, reaching a peak in December. This increase in androstenedione concentration correlated with the period of heavy accumulation of adipose tissue and increase in body mass. Antral follicles grow slowly during the period of high circulating androstenedione concentration from October to December. There was a sharp decline in androstenedione concentration during January. Simultaneously with the decline in androstenedione concentration, a sharp increase in size of the antral follicle and circulating estradiol concentration was noticed. Soon thereafter, one of the follicles ruptures, followed by fertilization and the commencement of the first pregnancy. During the second pregnancy, antral follicles first appeared during late pregnancy in March in the contralateral ovary that lacked the corpus luteum. One follicle developed rapidly and quickly attained a preovulatory stage in April. This is reflected in a sharp increase in estradiol concentration during this period. Ovulation was observed immediately following the first pregnancy in May. During this period, androstenedione concentration remained low. The results of the present study suggest that high androstenedione concentration during October to December (winter dormancy) may be responsible for slow follicular development and delays ovulation in T. longimanus. It is further hypothesized that the geographical variation in reproductive pattern of T. longimanus could be due to variation in the duration of fat storage and associated changes in the androstenedione concentration.

Adaptation, Physiological↗

Light flashes of different durations (0.063-3.33 msec) phase shift the circadian flight activity of a bat.

The phase-response curve (PRC) for the circadian rhythm in the flight activity of a cave-dwelling bat, Hipposideros speoris, constructed with 0.063-msec light flashes, reported here is the first of its kind for any circadian system and is unlike any other phase-response curves constructed for other nocturnal animals. The phase responding with maximal advances (90 degrees) and the phase responding with maximal delays (0 degree) of this PRC were exposed to light flashes of systematically varying durations from 0.083 to 3.33-msec. For 0 degree phase, the flashes of 0.063-3.33 msec effected delay phase shifts of comparable magnitude. For 90 degrees phase, the flashes of 0.063-1.0 msec effected advance phase shifts, whereas 3.33-msec flashes effected unmistakable delay phase shifts with advancing transients. Phase shifts evoked with such light flashes are further compared with phase shifts evoked with pulses of longer durations (15 min to 2.8 hr) for H. speoris.

Animals↗

Histology and histochemistry of the parotid and the principal and accessory submandibular glands of the little brown bat.

The parotid and the principal and accessory submandibular glands of the little brown bat. Myotis lucifugus (Vespertilionidae), were examined using light microscopy and staining methods for mucosubstances. The parotid gland is a compound tubuloacinar seromucous gland. Parotid gland secretory cells contain both neutral and nonsulfated acidic mucosubstances. The principal and accessory submandibular glands are compound tubuloacinar mucus-secreting glands. They contain somewhat atypical mucus-secreting demilunar cells that often appear to be interspersed between mucous tubule cells. The mucous tubule cells in both the principal and accessory submandibular glands contain sulfonmucins. Demilunar cells of the principal submandibular gland contain moderate amounts of nonsulfated acidic mucosubstances, but the corresponding cells of the accessory submandibular gland contain considerable neutral mucosubstance with very little acid mucosubstance. Intercalated ducts composed of cuboidal or low columnar epithelial cells are present in all three glands. Striated ducts in all glands are composed of columnar cells whose apices bulge into the ductal lumina. Excretory ducts are composed of simple columnar epithelium, with occasional basal cells that suggest a possible pseudostratified nature. The cells of the excretory ducts also have bulging apices. All duct types contain apical cytoplasmic secretory material that is a periodic acid-Schiff positive, neutral mucosubstance. Ductal apical secretory material is more evident in intercalated and striated ducts than in excretory ducts.

Animals↗

Histochemistry of flight muscles in the Jamaican fruit bat, Artibeus jamaicensis: implications for motor control.

Two fast-twitch fiber types are histochemically identified in the primary flight muscles of Artibeus jamaicensis. These are classified as type IIa and IIb according to an acid-preincubation staining protocol for myosin ATPase. All fibers in the bat flight muscles exhibit relatively intense staining properties for NADH-TR, suggesting a high oxidative capacity. The glycolytic potential of all fibers is rather low, as assessed by stains for alpha-GPD. This two-type histochemical profile appears to parallel biphasic electromyographic patterns observed in these muscles and leads us to propose that flight muscle histochemistry and activation are mediated by a "two-gear" neuromuscular control system. In contrast, earlier studies on Tadarida brasiliensis demonstrate the existence of a "one-gear" neuromuscular control system, exemplified by the presence of one fiber type. These observations are discussed with respect to the natural history and flight styles of several species.

Adenosine Triphosphatases↗

Morphology of the abdominal wall in the bat, Pteronotus parnellii (Microchiroptera: Mormoopidae): implications for biosonar vocalization.

We investigated the structure of the abdominal wall of Pteronotus parnellii and made comparisons with eight other species of Microchiroptera and one megachiropteran. Similar to other mammals, the abdominal wall of bats consists of the three flank muscles laterally and the m. rectus abdominis ventrally. In Microchiroptera, flank muscles are mostly confined to dorsal portions of the wall. The mm. transversus abdominis and obliquus internus abdominis form the bulk of the wall; the m. obliquus externus is poorly developed. Ventrolaterally, a large portion of the wall is a dense, bilaminar aponeurosis, composed of collagen, elastin, and fibroblasts. The thicker, superficial lamina derives from the mm. obliquus internus and transversus abdominis. The deep lamina is a continuation of the transversalis fascia. Collagen fibers of the two fused laminae are oriented orthogonally, resulting in a resilient, composite fabric. Fascicles of the flank muscles are oriented along the margins of the aponeurosis so that their forces appear to be concentrated onto the aponeurosis. We suggest that this system is adapted for the regulation and generation of intra-abdominal pressure. The abdominal wall of Pteropus, the one megachiropteran examined, lacks the derived aponeurosis and is similar to other mammals. We consider the abdominal wall of Microchiroptera to be analogous to the diaphragma, in that it functions in the regulation of pressure within body cavities and facilitates biosonar vocalization.

Abdominal Muscles↗

Fine structure of the neck of epididymal spermatozoa of Schreiber's long-fingered bat (Chiroptera: Mammalia).

The structure of the spermatozoon of the long-fingered bat (Miniopterus schreibersii) was studied using transmission electron microscopy. The neck is characterized by two asymmetrically arranged features that allow the orientation of sections in different planes and therefore aid the interpretation of the structure. There are six segmented columns, two major and four minor, in the anterior 79% of the neck, and it is only posterior to this that the major columns split. One major column is associated with doublets 9, 1, and 2, and the other major column, with doublets 5 and 6. The proximal centriole is surrounded by the capitulum and segmented columns, and the distal centriole persists and is apparent in both longitudinal and transverse sections. The central pair of microtubules pass through the distal centriole and terminate at the proximal centriole. A line drawn through the two central microtubules divides the head into left and right halves. Development of the neck begins during early spermiogenesis at which time the two centrioles are associated with the production of the capitulum, segmented columns, and axoneme complex.

Animals↗

Ontogeny of vocal signals in the little brown bat, Myotis lucifugus

This study reports developmental changes in the vocal signals produced by wild-caught pre-volant and volant Myotis lucifugusAudio recordings were made from young animals (1-33 days old) and adults (over 1 year of age). The animals were removed from an attic maternity roost and studied individually in a room below. To stimulate flight-related behaviours, animals were released from a launching platform via a hinged floor, and their vocalizations were recorded as they approached a soft foam pad below. When the hinged floor opened, the youngest animals (1-4 days old) typically dropped onto the pad, but older animals either flapped their wings to achieve some horizontal displacement (>4 days) or sustained horizontal flight (>17 days). Vocalizations recorded under these conditions showed frequency modulation characteristic of adult echolocation sounds, even in animals as young as 4 days. Ontogenetic trends showed an increase in sound frequency, an increase in sound repetition rate and a decrease in sound duration as the animals matured. These data are discussed in the context of the development of echolocation behaviour in bats.

Journal Article↗